LOCUS       Exported                7236 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  MAC-tagged gene expression.
ACCESSION   .
VERSION     .
KEYWORDS    MAC_C_SCO1
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7236)
  AUTHORS   Liu X, Salokas K, Tamene F, Jiu Y, Weldatsadik RG, Ohman T, 
            Varjosalo M
  TITLE     An AP-MS- and BioID-compatible MAC-tag enables comprehensive mapping
            of protein interactions and subcellular localizations.
  JOURNAL   Nat Commun. 2018 Mar 22;9(1):1188. doi: 10.1038/s41467-018-03523-2.
  PUBMED    29568061
REFERENCE   2  (bases 1 to 7236)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7236
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        9..388
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        389..592
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     543..563
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    594..612
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    615..633
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     643..667
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    810..834
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(1747..1771)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1818..1841
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             1878..1901
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             1932..1958
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1959..2918
                     /codon_start=1
                     /product="promiscuous R118G mutant of the E. coli biotin 
                     protein ligase (Cronan, 2005)"
                     /label=BirA(R118G)
                     /note="also known as BirA* or BioID"
                     /translation="KDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRDW
                     GVDVFTVPGKGYSLPEPIQLLNAKQILGQLDGGSVAVLPVIDSTNQYLLDRIGELKSGD
                     ACIAEYQQAGRGGRGRKWFSPFGANLYLSMFWRLEQGPAAAIGLSLVIGIVMAEVLRKL
                     GADKVRVKWPNDLYLQDRKLAGILVELTGKTGDAAQIVIGAGINMAMRRVEESVVNQGW
                     ITLQEAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIGD
                     KEIFGISRGIDKQGALLLEQDGIIKPWMGGEISLRSAEK"
     primer_bind     complement(2962..2979)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2968..3192
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3325..3344)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    3476..3523
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             3531..4553
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     polyA_signal    4683..4804
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4720..4739)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4774..4793
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4853..4869)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4853..4869)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4866..4888)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4877..4893
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(4901..4931)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4946..4967
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5084..5101)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5255..5843)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5335..5354)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6014..6874)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6637..6656
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6875..6979)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7054..7073)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 tacgcgttga cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt
       61 tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg
      121 accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc
      181 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc
      241 agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg
      301 gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat
      361 ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg
      421 tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag
      481 tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt
      541 gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctctccctat
      601 cagtgataga gatctcccta tcagtgatag agatcgtcga cgagctcgtt tagtgaaccg
      661 tcagatcgcc tggagacgcc atccacgctg ttttgacctc catagaagac accgggaccg
      721 atccagcctc cggactctag cgtttaaact taagcttggt accgagctcg gatccactag
      781 tccagtgtgg tggaattctg cagatatcaa caagtttgta caaaaaagca ggctccacca
      841 tggcgatgct ggtcctagta cccggacgag ttatgcggcc tctgggtggc caactttggc
      901 gcttcttgcc tcgcggactc gagttttggg gcccagccga ggggactgcg agagtcttgc
      961 tgaggcagtt ctgcgcgcgg caagcggagg cgtggcgtgc ctcggggcgc cctggctatt
     1021 gcctgggaac ccggcccctc agcactgcga ggccgccacc cccgtggtcg cagaagggcc
     1081 ccggagactc cacgcgcccc tcgaagcccg ggcctgtttc ctggaagtct ttagcaatca
     1141 catttgctat tggaggagct ttactggctg gaatgaagca cgtcaagaaa gaaaaggcag
     1201 agaagttaga gaaggaacgg cagcgacaca tcggcaagcc tttacttggg ggaccgtttt
     1261 ccctcacaac tcatactggg gagcgtaaaa ctgacaagga ctacttgggt cagtggttat
     1321 tgatttattt tggcttcact cattgccctg atgtctgtcc agaagaacta gaaaagatga
     1381 ttcaagtcgt ggatgaaata gatagcatta caactctgcc agatctaact ccacttttca
     1441 tcagcattga cccagagagg gacacaaaag aagccatcgc aaattatgtg aaagaatttt
     1501 ctcccaaact ggttggcttg actggcacga gagaagaggt cgatcaagtg gccagagcat
     1561 acagagtgta ttacagccct ggccccaagg acgaagatga agactacata gtggatcaca
     1621 caataataat gtacttgatt ggaccagatg gtgagtttct agattatttt ggccagaaca
     1681 agaggaaggg agaaatagct gcttcaattg ccacacacat gaggccatac agaaaaaaga
     1741 gcttggaccc agctttcttg tacaaagtgg ttgatatcaa ggccgccgac atcaccagcc
     1801 tgtacaagaa agtggggtgg agccatcccc agttcgagaa gggcggagga tctggcggcg
     1861 gaagcggcgg aggaagctgg tcccaccctc agtttgagaa aggcaccgag ctgggcagca
     1921 ccatggccag ctacccctac gacgtgcccg actacgccaa ggacaacacc gtgcccctga
     1981 agctgatcgc cctgctggcc aacggcgagt tccactctgg cgagcagctg ggcgagacac
     2041 tgggcatgag cagagccgcc atcaacaagc acatccagac cctgcgggac tggggcgtgg
     2101 acgtgttcac agtgcccggc aagggctaca gcctgcccga gcccatccag ctgctgaacg
     2161 ccaagcagat cctgggccag ctggatggcg gatctgtggc cgtgctgccc gtgatcgaca
     2221 gcaccaacca gtatctgctc gaccggatcg gcgagctgaa gtccggcgac gcctgtatcg
     2281 ccgagtacca gcaggccggc agaggcggca ggggcagaaa gtggttcagc cccttcggcg
     2341 ccaacctgta cctgagcatg ttctggagac tggaacaggg cccagccgcc gctattggcc
     2401 tgagcctggt catcggcatc gtgatggccg aggtgctgcg gaagctgggc gccgacaaag
     2461 tgcgcgtgaa gtggcccaac gacctgtatc tgcaggaccg gaagctggcc ggcatcctgg
     2521 tggaactgac cggcaaaaca ggcgacgccg cccagatcgt gatcggcgct ggcatcaaca
     2581 tggccatgcg gcgggtggaa gagagcgtgg tcaaccaggg ctggatcacc ctgcaggaag
     2641 ccggcatcaa cctggaccgg aacaccctgg ccgccatgct gatcagagag ctgagagccg
     2701 ccctggaact gttcgagcag gaaggcctgg ccccctacct gagcagatgg gagaagctgg
     2761 acaacttcat caaccggccc gtgaaactga tcatcggcga caaagagatc ttcggcatca
     2821 gccggggcat cgacaagcag ggggccctgc tcctggaaca ggacggcatc atcaagccct
     2881 ggatgggcgg cgagatcagc ctgcggagcg ccgagaagtg atgactcgag tctagagggc
     2941 ccgtttaaac ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt
     3001 gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat
     3061 aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg
     3121 tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg
     3181 tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg
     3241 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     3301 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     3361 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     3421 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt acctagaagt
     3481 tcctattccg aagttcctat tctctagaaa gtataggaac ttccttggcc aaaaagcctg
     3541 aactcaccgc gacgtctgtc gagaagtttc tgatcgaaaa gttcgacagc gtctccgacc
     3601 tgatgcagct ctcggagggc gaagaatctc gtgctttcag cttcgatgta ggagggcgtg
     3661 gatatgtcct gcgggtaaat agctgcgccg atggtttcta caaagatcgt tatgtttatc
     3721 ggcactttgc atcggccgcg ctcccgattc cggaagtgct tgacattggg gaattcagcg
     3781 agagcctgac ctattgcatc tcccgccgtg cacagggtgt cacgttgcaa gacctgcctg
     3841 aaaccgaact gcccgctgtt ctgcagccgg tcgcggaggc catggatgcg atcgctgcgg
     3901 ccgatcttag ccagacgagc gggttcggcc cattcggacc gcaaggaatc ggtcaataca
     3961 ctacatggcg tgatttcata tgcgcgattg ctgatcccca tgtgtatcac tggcaaactg
     4021 tgatggacga caccgtcagt gcgtccgtcg cgcaggctct cgatgagctg atgctttggg
     4081 ccgaggactg ccccgaagtc cggcacctcg tgcacgcgga tttcggctcc aacaatgtcc
     4141 tgacggacaa tggccgcata acagcggtca ttgactggag cgaggcgatg ttcggggatt
     4201 cccaatacga ggtcgccaac atcttcttct ggaggccgtg gttggcttgt atggagcagc
     4261 agacgcgcta cttcgagcgg aggcatccgg agcttgcagg atcgccgcgg ctccgggcgt
     4321 atatgctccg cattggtctt gaccaactct atcagagctt ggttgacggc aatttcgatg
     4381 atgcagcttg ggcgcagggt cgatgcgacg caatcgtccg atccggagcc gggactgtcg
     4441 ggcgtacaca aatcgcccgc agaagcgcgg ccgtctggac cgatggctgt gtagaagtac
     4501 tcgccgatag tggaaaccga cgccccagca ctcgtccgag ggcaaaggaa tagcacgtac
     4561 tacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     4621 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     4681 ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     4741 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     4801 cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc
     4861 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     4921 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     4981 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     5041 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     5101 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     5161 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     5221 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     5281 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     5341 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     5401 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     5461 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     5521 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     5581 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     5641 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     5701 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     5761 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     5821 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     5881 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     5941 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     6001 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     6061 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     6121 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     6181 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     6241 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     6301 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     6361 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     6421 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     6481 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     6541 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     6601 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     6661 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     6721 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     6781 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     6841 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     6901 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     6961 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc gacggatcgg
     7021 gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
     7081 aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat
     7141 ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc ttagggttag
     7201 gcgttttgcg ctgcttcgcg atgtacgggc cagata
//
